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Biochemistry & Molecular Biology

机译:生物化学与分子生物学

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A maize RNA binding motif protein-48 (RBM48) is a master-gene critical in the development and viability of seeds in maize. Mutations in the RBM48 master-gene cause severe developmental defects and exhibit prolific growth upon tissue culture. In this research project, RNA extracted from the mutant RBM48 and the wild-type RBM48 gene are compared using next-generation RNA sequencing to generate a set of candidate genes that are expressed differentially in the mutant gene. Next, RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction) is used to validate differential gene expression by making cDNA primers from mutant and wild-type RNA, amplifying the results using PCR, and using gel electrophoresis to experimentally validate the identities of differentially expressed genes. Bioinformatics tools make connections between the genes of interest and their biological function. For example, tools such as BLAST are used to find homologous genes in maize and other species to help us understand the functions of the differentially expressed genes in RBM48. Bioinformatics tools can further identify possible gene mechanisms of action that may be responsible for prolific growth with the hope that insights into the development of cancer may become possible. The research is significant from agricultural perspectives regardless since maize is a vital US crop.
机译:玉米RNA结合基序蛋白48(RBM48)是一个主要基因,对玉米种子的发育和生存能力至关重要。 RBM48主基因中的突变会导致严重的发育缺陷,并在组织培养后表现出丰富的生长。在该研究项目中,使用下一代RNA测序对从突变RBM48和野生型RBM48基因提取的RNA进行比较,以生成一组在突变基因中差异表达的候选基因。接下来,使用RT-PCR(逆转录酶-聚合酶链反应)通过从突变型和野生型RNA制备cDNA引物,使用PCR扩增结果,并使用凝胶电泳通过实验验证差异表达的身份,来验证差异基因的表达。基因。生物信息学工具在目标基因及其生物学功能之间建立联系。例如,使用诸如BLAST之类的工具来查找玉米和其他物种中的同源基因,以帮助我们了解RBM48中差异表达基因的功能。生物信息学工具可以进一步确定可能导致多产生长的可能的基因作用机制,并希望有可能深入了解癌症的发展。从玉米的角度来看,这项研究意义重大,因为玉米是美国至关重要的农作物。

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    《Michigan Academician》 |2018年第3期|365-373|共9页
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